LNCaP LNCaP E.V. LNCaP TG2 0 2 4 6 8 10 12 14 Fig. S1. Effect of TG2 expression on LNCaP cell growth (A) and Karyotyping of parental, vector-infected and TG2-transfected LNCaP cells (B) A B LNCaP LNCaP-EV LNCaP-TG2 Relative cell growth over 3 days
Jan 12, 2016
LNCaP LNCaP E.V.
LNCaP TG2
0
2
4
6
8
10
12
14
Fig. S1. Effect of TG2 expression on LNCaP cell growth (A) andKaryotyping of parental, vector-infected and TG2-transfected LNCaP cells (B)
A
B
LNCaP LNCaP-EV LNCaP-TG2
Rela
tive
cell
grow
th o
ver 3
day
s
Histones
p65
TG2
IκBα
LNCaP
Cont.
LNCaP
EV LNCaP
TG2
C N C N C N C N
PC3
Fig. S2. Co-localization of TG2 and NF-κB in nuclear fraction of prostate cancer cells (A). Transient down regulation of NF-kB partially restored AR expression in PC3 cells (B). Putative NF-κB binding sites in the promoter of AR gene (C and D).
Exon 1
Cont si
R
A
p65
Actin
p65 s
iR
AR
B
-38 bpGACCCGACTCGCAAACTGTTGCATTTGCTCTCCACCTCCCAGCGCCCCCTCCGAGATCCCGGGGAGCCAGCTTGCTGGGAGAGCGGGACGGTCCGGAGCAAGCCCAGAGGCAGAGGAGGCGACAGAGGGAAAAAGGGCCGAGCTAGCCGCTCCAGTGCTGTACAGGAGCCGAAGGGACGCACCACGCCAGCCCCAGCCCGGCTCCAGCGACAGCCAACGCCTCTTGCAGCGCGGCGGCTTCGA AGCCGCCGCCCGGAGCTGCCCTTTCCTCTTCGGTGAAGTTTTTAAAAGCTGCTAAAGACTCGG +246 bp
-760 bpGGGTGATTTTGCCTTTGAGAGTCTGGATGAGAAATGCATGGTTAAAGGCAATTCCAGACAGGAAGAAAGGCAGAGAAGAGGGTAGAAATGACCTCTGATTCTTGGGGCTGAGGGTTCCTAGAGCAAATGGCACAATGCCACGAGGCCCGATCTATCCCTATGACGGAATCTAAGGTTTCAGCAAGTATCTGCTGGCTTGGTCATG -460 bp
C
D
Fig. S3. TG2 and AR protein expression in selected prostate tumor samples (A). TG2 expression Is associated with increased accumulation of HIF-1α protein in prostate cancer cells (B)
TG2 AR
LnCa
p-EV
LnCa
p-TG2
PC3
HIF-1α
TG2
β-actin
A
B
Gene designation Primer Sequence (5’-3’)
AR F: TTGGATGGCTCCAAATCACR: GCAATGATACGATCG AGTTCC
E-cadherin F: TCCCATCAGCTGCCCAGA A A
R:TGACTCCTGTGTTCCTGT TA
N-cadherin F: ACAGTGGCCACCTACAAAGGR: CCGAGATGGGGTTGATAATG
Vimentin F: GAG AACTTTGCCGTTGAA GC
R: GCTTCCTGTAGGTGGCAATC
Fibronectin F: CAGTGGGAGACCTCGAGAAG R:TCCCTCGGAACATCAGAAAC
ZEB1 F: CTGAAGAGGACCAGAGGCAG
R: CCCAGAACTGCGTCACATGTC
ZEB2 F: AAGTACCGCCACGAGAAGAA
R: TTGTTTCTCATTCGGCCATT
Snail F: TTCTTCTGCGCTACTGCTGCGR:
GGGCAGGTATGGAGAGGAAGA
TG2 F: TATGGCCAGTGCTGGGTCTTCGCC R: GGCTCCAGGGTTAGGTTGAGCAGG
GAPDH F: ACCACAGTCCATGCCATCAC
R: TCCACCACCCTGTTGCTGTA
Table S1 – Primers used to amplify various transcripts